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Surface Energetice
Surface Energetice
ABSTRACT
This study investigated the interactive effects of Hepatitis C virus on human cells using the
contact angle approach. The methodology involves the use of sessile drop approach to de-
termine the contact angle formed on the infected and uninfected blood cells in the presences
of glycerin as the probe liquid. It was observed that the presence of the virus in the human
blood cells depleted the immune system of infected cells giving rise to a decreased CD4
count on the average of 514.5 ± 243.10 when compared with the uninfected cells CD4 count
of 1267.2 ± 368.27. The measurement of contact angle also unveils that among the blood
components separated in the course of the experiment, the white blood cell is the principal
target of the virus with the highest average contact angle of 63.4 ± 3.20 while the uninfected
white blood cells have a lower contact angle of 48.5 ± 2.75. The result of the measured con-
tact angle was used for MATLAB computation to determine the surface energy, force of
adhesion and the Hamaker coefficient. Response surface methodology was also employed in
this study to visualize the viral impact on the blood cells as well as generating model equa-
tions for prediction of the interaction between the virus and the blood cells. Infected sur-
faces on the average have higher values of Hamaker coefficient than uninfected surfaces. It
was discovered that an increase in the contact angles causes a significant increase in Ha-
maker coefficient with a corresponding decrease in the CD4 counts on the infected surfaces.
This increase is attributed to the presence of the HCV virus in the infected samples and the
highest value was observed in the white blood cell component. Computation of the com-
bined negative Hamaker coefficient revealed that there exists a possibility of separating the
1. INTRODUCTION
Hepatitis C is a global health burden to both industrialized and developing countries infecting an es-
timated 3% of the world population. It represents a viral pandemic that is 4 - 5 times more prevalent than
HIV infection, therefore the reduction of global mortality and morbidity related to hepatitis C is of great
concern to public health [1]. It is an infectious disease affecting the liver with no symptoms at the initial
stage of the infection and at long term impact of chronic infection causes permanent liver damage, cirrho-
sis, hepatocellular carcinoma and even death [2]. Despite infecting a great number of people worldwide,
the global burden of HCV infection remains at large a silent epidemic because acute infection is generally
asymptomatic and morbidity and mortality arise only after years of infection [3]. As at 2018, no approved
vaccine protects against hepatitis C virus [4], prevention includes testing of donated blood, harm reduc-
tion efforts among people who use intravenous drugs. However enormous advances have been made in the
treatment of hepatitis C virus infection, currently the combination of pegylated interferon-alpha and riba-
virin (RBV) is the standard treatment for the infection and it has so far achieved less than 50% success
with each HCV genotype showing differences in sustained virological response (SVR) to therapeutic
strategy, especially genotype 1 regarded as more problematic showing less than 50% HCV clearance [5].
A11 < A33 < A22 or A11 > A33 > A22 (1.6)
In relation to this study, A11 , A22 and A33 are Hamaker constants for uninfected white blood cell,
infected lymphocyte and serum respectively [10].
The experimental procedure for measuring contact angle and its interpretation in terms of young’s
equation was analyzed, stressing the need of how efficiently contact angle can be measured with strict ad-
herence [15].
2.2. Methods
2.2.1. Cluster of Differential Cell Count (CD4) of Blood Samples
Before the beginning of the experiment, the machine was cleaned by inserting into it 1600 μl each of
cleaning solution, decontamination solution and shealth fluid. 850 μl of count check beads green was used
on the machine to check for quality control. Quality of 25,100 per ml was obtained which falls between
standard acceptable ranges of 23,270 10% per ml. 20 μl of CD mAb PE monoclonal antibody was in-
troduced into a sample tube and 20 μl of EDTA whole blood was added to the tube and both were gently
mixed together. The mixture was incubated for 15 minutes in a dark field at room temperature. Added to
the mixture is 800 μl of no lysed buffer solution, mixed gently and ran the program. The CD4 cell count on
the whole blood samples was obtained using a Partec Cyflow Counter Machine. This helped to determine
the level and progression of the HCV depletion of the immune system of the patients. The machine dis-
plays automatically the number of CD4 + T cells.
Blood Samples Whole Blood (θ ˚) White Blood Cells (θ ˚) Red Blood Cells (θ ˚) Serum (θ ˚)
1 60 65 59 60
2 55 61 64 62
3 58 63 58 63
4 60 64 57 61
5 60 67 56 59
6 56 66 62 64
7 55 58 63 58
8 58 68 61 65
9 54 62 60 66
10 55 60 64 57
Average 57.1 63.4 60.4 61.5
SD 2.3781 3.2045 2.8752 3.0277
Blood Samples Whole Blood (θ ˚) White Blood Cells (θ ˚) Red Blood Cells (θ ˚) Serum (θ ˚)
1 50 47 48 58
2 51 46 57 55
3 40 48 45 45
4 50 52 50 56
5 45 50 52 53
6 56 51 51 57
7 55 45 49 54
8 50 49 54 52
9 43 44 47 50
10 55 51 53 51
Average 49.5 48.5 50.6 53.1
SD 5.3593 2.7508 3.5653 3.8427
The “Pred R-Squared” of 0.7290 is in reasonable agreement with the “Adj R-Squared” of 0.8987; i.e.
the difference is less than 0.2. “Adeq Precision” measures the signal to noise ratio. A ratio greater than 4 is
desirable. Table 6 predicts a ratio of 12.703 which indicates an adequate signal. This means that the model
is a good one for predicting Hamaker coefficient in relation with contact angle and interfacial energy due
to HCV infection.
A132 =2.05 − 0.021A + 0.12 B + 0.12 AB + 0.20 A2
(1.13)
+ 0.033B 2 + 0.20 A2 + 0.033B 2
The ANOVA indicates the equation and actual relationship between the response and significant va-
riables represented by the Equation (1.13) is accurate. The R2 value of 0.9409 indicates a good measure that
outcomes are likely to be predicted well by the developed models. The contour and the surface plot on
Figure 2 and Figure 3 also reveal graphically the interaction between the independent variable and the
response.
4. CONCLUSION
The measured contact angle data revealed that HCV infected surfaces have higher average contact
angles for all blood components than the uninfected sample. In the case of white blood cell, the contact
Figure 3. 3D surface plot of Hamaker coefficient for infected white blood cells.
A11 (mJ/m2) A22 (mJ/m2) A33 (mJ/m2) A131 (mJ/m2) A232 (mJ/m2) A132 (mJ/m2)
Infected 0.224 × 10−16 0.211 × 10−16 - 1.103 × 10−18 −0.150 × 10−18
Uninfected 0.125 × 10−16 - 0.160 × 10−16 0.21 × 10−18 - -
angle of the infected cell is 63.4˚ ± 3.20˚ and uninfected cell is 48.5˚ ± 2.75˚. The presence of the virus in
the infected sample causes the surfaces to be poorly wetted and as such leads to an increase in the contact
angle. The infected samples subjected to various treatments were observed to exhibit a reduction in meas-
ured contact angle. The negative value of the combined Hamaker coefficient (A132) for HCV infected sam-
ple given as −0.150 × 10−25 J signifies van der Waal repulsion in the interacting particles indicating that the
virus can be isolated from the lymphocyte when the serum acting as the intervening medium is altered. All
the drugs used have a reduction effect on the surface energy of the serum (A33) and an increasing effect on
the surface energy of the infected lymphocyte (A22).
DECLARATION
Ethical clearance was dully obtained from the relevant authorizing office at Chukwuemeka Odu-
megwu Ojukwu Teaching Hospital Amaku before this research was conducted.
CONFLICTS OF INTEREST
The authors declare no conflicts of interest regarding the publication of this paper.
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ABBREVATIONS
Cluster of Differentiation 4 (CD4), Human immunodeficiency virus (HIV), Hepatitis c virus (HCV), Ri-
bonucleic Acid (RNA), Combined Negative Hamaker coefficient (A132)